Novel brassica plants

ABSTRACT

The present invention relates to novel  brassica  plants, in particular to novel cauliflower plants. In one embodiment, the novel cauliflower plants provided herein comprise a long stem and are suitable for mechanical harvesting. The application also further discloses seeds the cauliflower plants of the present invention and parts thereof, for example pollen, ovules and curds. The application also further discloses methods of using a plant of instant invention, such as methods of producing a cauliflower curd of the instant invention, and methods of harvesting the curds of plants of the instant invention.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.60/687,768, filed Jun. 6, 2005. The above application is incorporatedherein by reference in its entirety.

FIELD OF THE INVENTION

The present invention relates to novel brassica plants, in particular tonovel cauliflower plants. The present invention also relates to seedsand parts of said plants, for example curds. The present invention alsorelates to methods of making and using such plants.

BACKGROUND OF THE INVENTION

Cauliflower (Brassica oleracea var. botrytis) is a member of theCruciferae (mustard) family and is an important crop grown worldwide.The world production of cauliflower is close to 5 millions tons a year,covering an area of around 300,000-400,000 hectares. The edible part ofthe plant is the head, also called curd, which comprises of the arrestedinflorescence and floral meristems. Cauliflower is an appreciatedvegetable because of its pleasant taste and nutritive value. The percapita consumption of fresh cauliflower in the United States wasestimated at 0.73 kg in 1982 (see e.g. Ib Libner Nonnecke, VegetableProduction, 1989, Van Nostrand Reinhold, NY Publisher, pp. 382-394).However, cauliflower is a difficult crop to grow. It has very preciseclimatic requirements, which, if ignored, cause failure or partialfailure under field conditions.

The cauliflower curd is very delicate and easily damaged, and isexclusively harvested by hand. This is however inefficient and costly,and it is estimated that 30-50% of the production costs for cauliflowerare harvesting costs. Attempts have been made to harvest cauliflowercurds mechanically, but they have faced difficulties in identifying themature curds and detaching them from the plant without damaging thecurds. Recently, a mechanical harvester specifically designed forcauliflower was described. This mechanical harvester uses sensortechnology and image processing (see HortLINK Caulicut project,http://www.ost.gov.uk/link/linkcasestudies/cauliflowersapril03.pdf).However, the cost of the machines was reported to be high. This wouldrequire a substantial upfront investment, and it remains to be seenwhether the prototypes described will be commercially successful.

There is therefore an unfulfilled need to reduce costs of production forcauliflower. In particular, there is an unfulfilled need fordevelopments allowing increased efficiency and reductions in the costsof harvesting marketable cauliflower curds.

SUMMARY OF THE INVENTION

The instant application addresses the unfulfilled needs to increase theefficiency and to reduce the production costs for cauliflower, inparticular to reduce costs of harvesting of cauliflower curds. Toaddress these needs, the instant invention provides novel cauliflowerplants with a special stem structure, especially cauliflower plantscomprising a long stem. In one embodiment, the cauliflower plants of thepresent invention are suitable for mechanical harvest.

In one embodiment, the stem of a plant of the present invention islonger than that of currently existing cauliflower plants havingcommercially desirable attributes, such as capable of producing amarketable curd, and requiring approximately the same number of growingdays to reach maturity. In one embodiment, the stem of a plant of thepresent invention is longer than that of currently existing cauliflowerplants in the same cycle having commercially desirable attributes, suchas capable of producing a marketable curd.

The feature of elongated stems offers the advantage of providing enoughroom between the ground and the bottom of the curd and thus facilitatesthe harvest of the curds. The feature of elongated stems also allowseconomical and efficient mechanical harvesting of the curd, whileminimizing the risk of damage by miscutting and soil contamination ofthe curd. This is particularly useful for cauliflower plants of shortercycles, which tend to grow shorter stems. Accordingly, in oneembodiment, a cauliflower plant of the instant invention is a plant ofthe short cycle or medium cycle.

In one embodiment, a plant of the instant invention further comprises atrait of loose foliage. In one aspect, the total leaf weight of a plantof the instant invention is lower than the total leaf weight ofcurrently existing cauliflower plants having commercially desirableattributes, such as capable of producing a marketable curd. In anotheraspect, the leaf volume index of a plant of the instant invention issmaller than the leaf volume index of currently existing cauliflowerplants having commercially desirable attributes, such as capable ofproducing a marketable curd. Plants with loose foliage allow for faster,more convenient and more efficient harvesting of the curds, and reducethe risk of jamming a mechanical harvester during harvest.

In one embodiment, the average internode length of the stem of a plantof the present invention is longer than that of currently existingcauliflower plants having commercially desirable attributes, such ascapable of producing a marketable curd. The feature of longer internodesoffers the advantage of providing longer stems while maintaining a lownumber of leaves on the stem or reducing the number of leaves on thestem.

In one embodiment, a plant of the instant invention further comprises atrait of a persistent white curd. The trait of persistent white curdprevents or slows down cauliflower curds from turning yellow whenexposed to sunlight, in particular in plants with curds that are notcovered or only partially covered by inner leaves, for example plantswith a loose foliage.

In one embodiment, a plant of the instant invention is a cauliflowerplant having commercially desirable attributes. In one embodiment, aplant of the instant invention is a cauliflower plant is capable ofproducing a marketable curd. In one embodiment, the curd of acauliflower plant of the present invention is used for fresh marketconsumption and in the processing industry.

In one embodiment, a plant of the present invention is an inbred line ora hybrid. In one embodiment, a plant of the instant invention is adihaploid. In one embodiment, an inbred line of the present invention isa dihaploid. In one embodiment, at least one of the parents of a hybridof the instant invention is a dihaploid. Dihaploid plants allow for highlevels of uniformity of the plants of the instant invention. In oneembodiment, a plant of the instant invention is capable of producing acurd that can be commercialized on the market.

In one embodiment, a plant of the present invention comprises anycombination of the traits of long stem, long internodes, number ofinternodes, loose foliage, low total leaf weight, small leaf volumeindex, and persistent white curd, and short or medium cycle anduniformity, as described herein. In one embodiment, a plant of thepresent invention comprises a long stem, loose foliage and persistentwhite curd, as described herein. In one embodiment, such plant is of theshort cycle or medium cycle.

The application further discloses seeds the cauliflower plants of thepresent invention and parts thereof, for example cells, pollen, ovulesand curds. The present application also further discloses methods ofusing a plant of instant invention, such as methods of producing acauliflower curd of the instant invention, and methods of harvesting thecurds of plants of the instant invention. The present application alsofurther discloses methods of producing seeds of a cauliflower plant ofthe instant invention, and methods of vegetatively propagating acauliflower plant of the instant invention.

In one embodiment, a plant of the instant invention is obtained bycrossing a cauliflower plant with a broccoli plant, and selecting aplant comprising a long stem. Accordingly, in one embodiment, theinstant application also provides methods of producing a cauliflowerplant of the present invention comprising crossing a cauliflower plantwith a broccoli plant and selecting for plants comprising a long stem.In one embodiment, the method further comprises selecting forcharacteristics of a cauliflower curd. In one embodiment, the instantinvention further provides plants obtained by such methods.

In one embodiment, the instant application also provides methods forproducing a cauliflower plant comprising a long stem comprising crossinga cauliflower plant and a plant of line 03P001, representative seed ofwhich being deposited under accession number NCIMB 41228, and selectinga plant comprising a long stem. In one embodiment, the instant inventionfurther provides cauliflower plants obtained by such methods.

The plants of the present invention make the harvest of cauliflowercurds by hand easier and faster, and thus more efficient. In oneembodiment, the plants of the present invention also allow mechanicalharvest of the curds using fully automatic or semi-automatic harvesters.Simple and inexpensive machinery can be used with the plants of thepresent invention, while keeping losses caused by miscutting andcontamination with soil particles to a minimum. Harvest costs forcauliflower curds, and thus production costs for cauliflower can beaccordingly reduced.

The invention therefore provides:

A cauliflower plant comprising at maturity a stem with an averageinternode length of at least 1.25 cm. In one embodiment, the averageinternode length is at least 1.30 cm, at least 1.35 cm or at least 1.40cm. In one embodiment, the average internode length is 1.25 cm to 2.00cm. In one embodiment, the stem comprises 18 internodes to 26internodes, 19 internodes to 25 internodes, or 20 internodes to 24internodes. In one embodiment, the average internode length and thenumber of internodes are measured when the plant is grown underconditions representative for a continental climate, e.g. in Ocsa,Hungary, and planted during the 4^(th) week of May and transplantedduring the 5th week of June.

The invention further provides:

A cauliflower plant comprising at maturity a stem with an averageinternode length of at least 80% of the average internode length of aplant of line 03P001, representative seed of which being deposited underaccession number NCIMB 41228. In one embodiment, the average internodelength is at least 85% or at least 90% of the average internode lengthof a plant of line 03P001. In one embodiment, the average internodelength is 80% to 140% of the average internode length of a plant of aplant of line 03P001. In one embodiment, the number of internodes ofsaid plant is less than 120%, less than 115% or less than 110% of thenumber of internodes of a plant of line 03P001. In one embodiment, thenumber of internodes of said plant is 80% to 120% the number ofinternodes of a plant of line 03P001.

The invention further provides:

Any one of the cauliflower plants above, wherein the stem length of saidplant is at least 85% or at least 90% of the stem length of a plant ofline 03P001, representative seed of which being deposited underaccession number NCIMB 41228. In one embodiment, the stem length is 85%to 130% of the stem length of a plant of line 03P001.

The invention further provides:

Any one of the cauliflower plants above, wherein the plant is a plant ofthe short cycle or of the medium cycle.

The invention further provides:

A cauliflower plant comprising at maturity a stem with a length of atleast 22.5 cm, when said plant is grown under conditions representativefor a continental climate, e.g. in Ocsa, Hungary, and planted during the3^(rd) week of May and transplanted during the 3^(rd) week of June orwhen said plant is grown under conditions representative for aMediterrenean climate, e.g. in Murcia, Spain, and planted during the1^(st) week of August and transplanted during the 1^(st) week ofSeptember, wherein said cauliflower plant is a plant of the short cycleor of the medium cycle. In one embodiment, the length of said stem is atleast 25 cm. In one embodiment, the length of said stem is 22.5 cm to 35cm.

The invention further provides:

A cauliflower plant comprising at maturity a stem with a length of atleast 30 cm, when said plant is grown under conditions representativefor a continental climate, e.g. in Ocsa, Hungary, and planted during the4th week of May and transplanted during the 5th week of June, whereinsaid cauliflower plant is a plant of the short cycle or of the mediumcycle. In one embodiment, the length of said stem is at least 32.5 cm.In one embodiment, the length of said stem is 30 cm to 45 cm.

The invention further provides:

Any one of the cauliflower plants above, wherein a stem of said plantcomprises 18 internodes to 26 internodes, 19 internodes to 25internodes, or 20 internodes to 24 internodes.

The invention further provides:

A cauliflower plant comprising at maturity a stem with a length of atleast 85% of the stem length of a plant of line 03P001 , representativeseed of which being deposited under accession number NCIMB 41228,wherein said plant is a plant of the short cycle or of the medium cycle.In one embodiment, the stem is at least 90% of the length of a stem of aplant of line 03P00 1. In one embodiment, the length of said stem is 85%to 130% of the length of a stem of a plant of line 03P001.

The invention further provides:

Any one of the cauliflower plants above, wherein the number ofinternodes of said plant is less than 120%, less than 115% or less than110% of the number of internodes of a plant of line 03P001,representative seed of which being deposited under accession numberNCIMB 41228. In one embodiment, the number of internodes of said plantis 80% to 120% the number of internodes of a plant of line 03P001.

The invention further provides:

Any one of the cauliflower plants above, wherein the total leaf weightof said plant is less than 150%, less than 140%, less than 130%, lessthan 120% or less than 110% of the total leaf weight of a plant of line03P001, representative seed of which being deposited under accessionnumber NCIMB 41228. In one embodiment, the total leaf weight of saidplant is 80% to 150% of the total leaf weight of a plant of line 03P001.

The invention further provides:

Any one of the cauliflower plants above, wherein the leaf volume indexof said plant is less than 130%, less than 125, less than 120% or lessthan 110% of the leaf volume index of a plant of line 03P001,representative seed of which being deposited under accession numberNCIMB 41228. In one embodiment, the leaf volume index of said plant is80% to 130% of the leaf volume index of a plant of line 03P001.

The invention further provides:

Any one of the cauliflower plants above, wherein the plant is an inbredline, a hybrid or a dihaploid. In one embodiment, the plant ismale-sterile. In one embodiment, the plant is obtainable by crossing abroccoli plant with a cauliflower plant.

The invention further provides:

Any one of the cauliflower plants above, wherein the plant comprises thegenetic background of line 03P001, representative seed of which beingdeposited under accession number NCIMB 41228. A cauliflower plantexhibiting a stem structure as described above, wherein the traitresponsible for said stem structure is obtainable through introgressionfrom a predecessor plant, which contains the genetic make-up of line03P00 1, representative seed of which being deposited under accessionnumber NCIMB 41228. A cauliflower plant exhibiting a stem structure asdescribed above, wherein the trait responsible for said stem structureis introgressed form a predecessor plant, which contains the geneticmake-up of line 03P001, representative seed of which being depositedunder accession number NCIMB 41228.

The invention further provides:

Seed of any one of the cauliflower plants above.

Part of any one of the cauliflower plants above. In one embodiment, thepart is a cell, pollen, an ovule, or a curd.

The invention further provides:

A method of producing a cauliflower curd comprising growing any one ofthe cauliflower plants above until a curd is produced and harvestingsaid curd. An agronomic method comprising growing any one of thecauliflower plants above until a curd ready for harvesting is produced;harvesting the curd. In one embodiment, the curd is harvestedmechanically.

The invention further provides:

A method of harvesting a cauliflower curd comprising growing any one ofthe cauliflower plants above until a curd is produced and mechanicallyharvesting said curd. In one embodiment, a blade is used in said step ofmechanically harvesting said curd.

The invention further provides:

A method of producing seed of a cauliflower plant comprising growing anyone of the cauliflower plants above; allowing the cauliflower plant toself-pollinate; and harvesting seeds from the cauliflower plant.

The invention further provides:

A method of vegetatively propagating a cauliflower plant comprisingcollecting a tissue of any one of the cauliflower plants above;cultivating the tissue to obtain proliferated shoots; and rooting theproliferated shoots to obtain rooted plantlets.

The invention further provides:

A method for producing a plant comprising a long stem comprisingcrossing a broccoli plant and a cauliflower plant; obtaining a progenyplant from the cross; measuring the stem length or average internodelength of the progeny plant; and selecting a plant comprising a longstem or long average internode length as described herein. In oneembodiment, the method further comprises obtaining a dihaploid of theplant selected above.

A plant obtainable by any one of the methods above.

The invention further provides:

A method for producing a cauliflower plant comprising a long stemcomprising crossing a cauliflower plant and a plant of line 03P001,representative seed of which being deposited under accession numberNCIMB 41228; obtaining a progeny plant from the cross; measuring thestem length or average internode length of the progeny plant; andselecting a cauliflower plant with a long stem or long average internodelength as described herein.

A cauliflower plant obtainable by any one of the methods above.

DEFINITIONS

Growing days: refers to the number of days from the date oftransplanting of young plants into a field or a plot to the date when50% of the plants in the field or plot reach maturity.

Maturity: a cauliflower plant has reached maturity when it has produceda curd that can be harvested and commercialized on the market (i.e. amarketable curd). Based on the certain market preference, the curd sizecan range from 15 cm to 30 cm in diameter.

Cycle: number of growing days under particular growing conditions. Thelength of the cycle is based on the genetics of a cauliflower plant anddepends on the climatic conditions, in particular the night temperature,under which the plant is grown. The cycle is generally defined forspecific climatic conditions, locations, sowing and transplanting times,or a combination thereof. Cauliflower plants, such as plants of inbredlines or hybrids, are for example grouped into short cycle (SC), mediumcycle (MC), medium-long (MLC), long cycle (LC) and very long cycle(VLC).

Short cycle (SC): 60-70 growing days after transplanting, based on aJanuary sowing and March transplanting in the conditions ofNorth-Western Europe, for example Enkhuizen, The Netherlands. Typicalplants of the SC are for example plants of variety Vinson (Seminis).

Medium cycle (MC): 60-72 growing days after transplanting, based on aMay sowing and June transplanting in the conditions of North-WesternEurope, for example Enkhuizen, The Netherlands. Typical plants of the MCare for example plants of variety Fremont (Seminis), Tetris (SyngentaSeeds) or Lecanu (Syngenta Seeds).

Medium long cycle (MLC): 73-90 growing days after transplanting, basedon a May sowing and June transplanting in the conditions ofNorth-Western Europe, for example Enkhuizen, The Netherlands. Typicalplants of the MLC are for example plants of variety Amerigo (SyngentaSeeds).

Long cycle (LC): Typical plants of the LC are for example plants ofvariety Belot (Bejo). Very long cycle (VLC): Typical plants of the VLCare for example plants of variety Mayfair (Syngenta Seeds).

Loose foliage: Reduced amount of foliage due to either smaller number ofleaves, reduced leaf size, or a combination thereof. Loose foliage isfor example measured by the leaf volume index or total leaf weight.Loose foliage may also be measured by the number of internodes on thestem of a plant.

Persistent white curd: This trait prevents or slows down cauliflowercurd turning yellow after exposure to sunlight. (see for example DicksonM. H. and Lee C. Y. Persitent white curd and other curd characters ofthe cauliflower. 1980 J. Amer. Soc. Hort. Sci. 105(4):533-535).

Trait: characteristic or phenotype. In the context of the presentinvention a trait is for example a long stem, long internodes, loosefoliage or persistent white curd, as described herein. A trait may beinherited in a dominant or recessive manner, or in a partial orincomplete-dominant manner. A trait may be monogenic or polygenic, ormay also result from the interaction of one or more genes with theenvironment.

Monogenic: determined by a single locus.

Polygenic: determined by more than one locus.

Dominant: results in a complete phenotypic manifestation at heterozygousor homozygous state.

Recessive: manifests itself only when present at homozygous state.

Partial or incomplete-dominance: when present at the heterozygous stagedetermines a phenotype that is intermediate to that of the homozygousstage or when the trait is absent.

Locus: region on a chromosome, which comprises a gene contributing to atrait.

Genetic linkage: association of characters in inheritance due tolocation of genes in proximity on the same chromosome. Measured bypercent recombination between loci (centi-Morgan, cM).

Isogenic: plants, which are genetically identical, except that they maydiffer by the presence or absence of a gene, a locus conferring a traitor heterologous DNA sequence.

Marker assisted selection: refers to the process of selecting a desiredtrait or desired traits in a plant or plants by detecting one or morenucleic acids from the plant, where the nucleic acid is associated withthe desired trait.

Dihaploid: doubling of haploid (single chromosome) status of the genome(e.g. through anther culture or microspore culture) giving a completehomozygous plant.

“Tester” plant: plant used to characterize genetically a trait in aplant to be tested. Typically, the plant to be tested is crossed with a“tester” plant and the segregation ratio of the trait in the progeny ofthe cross is scored.

Gene: Unit of inheritance. Genes are located at fixed loci inchromosomes and can exist in a series of alternative forms calledalleles.

Allele: One of a pair or series of forms of a gene, which arealternative in inheritance because they are situated at the same locusin homologous chromosomes.

Homozygous: Having like alleles at one or more corresponding loci onhomologous chromosomes.

Heterozygous: Having unlike alleles at one or more corresponding loci onhomologous chromosomes.

As used herein, the term “plant” includes plant cells, plantprotoplasts, plant cells of tissue culture from which cauliflower plantscan be regenerated, plant calli, plant clumps, and plant cells that areintact in plants or parts of plants, such as pollen, flowers, seeds,curds, stalks, stumps, leaves and the like.

Plant Height: measured in centimeters (cm) from the soil line to the topof the leaves.

Stem length: measured in centimeters from the soil line to the bottom ofthe curd.

Average internode length: Stem length at maturity divided by the numberof internodes on the stem.

Number of internodes: number of internodes on a stem at maturity.

Leaf Width: measured in centimeters at the widest part of the leaf blademidpoint of the plant including the petiole.

Leaf Length: length of leaf blade measured in centimeters from themidpoint of the plant including excluding the petiole.

Leaf volume index: leaf length in centimeters multiplied by leaf widthin centimeter, divided by 1,000.

Total leaf weight: weight of all the leaves on a plant at the time ofharvest of the curd.

Curd Diameter: measured at the widest diameter of the curd (fromoverhead) in centimeters.

Stem Diameter: measured in centimeters and is taken at a point justbelow the head.

Yield: the weight in grams for a harvested curd.

Color: means the color of the curd at maturity.

Field Holding Ability: means the ability of a plant to maintain goodcurd quality after the optimal harvest date.

Regeneration. Regeneration refers to the development of a plant fromtissue culture.

Single Gene Converted (Conversion). Single gene converted (conversion)plant refers to plants which are developed by a plant breeding techniquecalled backcrossing wherein essentially all of the desired morphologicaland physiological characteristics of a variety are recovered in additionto the single gene transferred into the variety via the backcrossingtechnique or via genetic engineering.

DETAILED DESCRIPTION OF THE INVENTION

The instant application addresses the need to reduce the productioncosts for cauliflower, in particular harvesting costs, and to allow formore efficient harvesting of marketable cauliflower curds. While effortshave been made to develop machinery specifically adapted to theharvesting of cauliflower curds, the inventors of the instantapplication have addressed the above needs by an alternative strategy,comprising developing novel cauliflower plants. In particular, theinventors of the instant application have addressed the above needs byproviding cauliflower plants with a special stem structure, especiallycauliflower plants comprising a long stem. In one embodiment, the plantsaccording to the instant invention are suitable for mechanicalharvesting.

Cauliflower is a cool season plant tolerating temperatures as low as 4°C. and as high as 38° C. Cauliflower seeds are typically sown in traysfilled with a soil mixture suitable for germination and raised to youngplants. Generally, after about 4-6 weeks the young plants aretransplanted to open fields. The plant density in a field may vary, butis typically around 3 plants per m². During the growing period plantsare watered either by drip irrigation or by sprinklers. If climate iswet enough no irrigation is needed.

Cauliflower plants initially grow in a vegetative phase followed by agenerative phase. Typically, the stems elongate during the vegetativephase, but their growth is generally reduced or even stops, as thegenerative phase begins. The generative phase is characterized by thedevelopment of the curd. Plants with early maturity (i.e. short cycle)have a short vegetative phase and need less growing days to produce acurd. They also tend to have short stems and produce fewer internodes.Plants with late maturity (i.e. longer cycle) have a longer vegetativephase and need more growing days. They also tend to produce longerstems, which is usually due to a higher number of internodes.Cauliflower plants need a certain “vernalization” (cool nights atapproximately 16-18° C.) to induce curd formation. Absent suchvernalization or curd induction, curd formation and consequentlymaturity is delayed. The duration of the vegetative phase depends on theclimatic conditions. The warmer the mean temperature, the more likely itis for the plant to remain in the vegetative cycle, which increases thetime needed to produce marketable curds. This typically results in moreinternodes and thus also more leaves. It is a general rule forcauliflower plants that the number of growing days increases when thenight temperature is high. Therefore, the number of growing daysdefining the cycle of a cauliflower plant is generally defined for acrop sowed and transplanted at a specific time (expressed in week, monthor season) in a particular climatic zone or location. The person skilledin the art would be able to recognize the cycle of a cauliflower plant.

Most cauliflower production is scheduled to accommodate specifictemperature requirements by utilizing cultivars that are adapted to thevariations of the seasons. Growers generally prefer plants with short tomedium-long cycles, because they occupy the fields for less time, thusallowing better land use and limiting the risk of damage to the crop dueto bad weather or disease. Also, in many climatic regions, plants withshorter maturity are desired, since they may otherwise be at risk ofhigh temperatures in the summer or frost in the late fall or winter.However, plants with shorter cycles tend to produce shorter stems, whichmakes them particularly ill-suited for mechanical harvesting, and whichmake hand harvesting particularly strenuous. The instant invention istherefore particularly beneficial to plants of the shorter cycles, suchas plants for the short to medium cycles, and, in one embodiment, aplant of the present invention is a plant of the short or medium cyclesas defined herein.

Cauliflower curds are exclusively harvested by hand. Therefore, thedevelopment of new cauliflower types and cultivars has focused on traitsfacilitating hand harvesting. Such traits are for example end-producttraits, such as curd quality and taste, and traits, which helped cropmaintenances, for example resistance to diseases. An important area ofresearch focused on developing cauliflower types with leaf structurescovering the curd to avoid the need to protect curd by hand during theharvest, and generally also characterized by heavy foliage. Anotherimportant area of research was to provide an upright plant habit toincrease the plant number per acreage.

By contrast, the features of long stem and long internodes areundesirable for cauliflower curds harvested by hand, because they makethe plants more vulnerable to lodging, which can cause rotting on thecurd or on the surrounding leaves. Lodged plants also make hand harvestmore difficult. The traits of long stem and long internodes weretherefore systematically eliminated from the cauliflower germplasm pool.

Accordingly, the inventors of the present invention have evaluated othersources for the traits of long stem and long internodes, in particularother brassica species. In one embodiment, the traits of long stem andlong internodes were obtained from another brassica species, in oneembodiment from broccoli (Brassica oleracea L. Italica). However,crosses between species can have unexpected, unfavorable consequences,in particular when the plant's architecture is modified. Also, thecauliflower curd is a very delicate structure with a particular taste,consistency and appearance, and other brassica species have numeroustraits, which are undesirable in a cauliflower curd. In particular, thehead type and the green color of the head of broccoli plants areundesired in cauliflower. Accordingly, in one embodiment, the trait oflong stem and long internodes according to the instant invention wasobtained from broccoli and successfully transferred to cauliflower,while surprisingly avoiding other undesired broccoli traits, and in oneembodiment, a cauliflower plant of the instant invention is acauliflower plant having commercially desirable attributes, such ascapable of producing a marketable curd.

In one embodiment, a cauliflower plant was crossed to a broccoli plant.The resulting F1 progeny was self-pollinated and selected for long stemand long internodes and for the best possible cauliflower curd. SelectedF2 progenies were again self-pollinated and selected for long stem andlong internodes and for the best possible cauliflower curd. In somecase, self-pollination and selection was repeated. The best F2 or F3plants were subjected to the dihaploid technique for line fixation inorder to get the highest possible uniformity of plants of a particularline. F1 and subsequent generations could also be back-crossed withcauliflower plants having desired characteristics to improve curdquality.

Accordingly, the instant application also provides methods of producinga plant of the present invention comprising crossing a cauliflower plantwith a broccoli plant and selecting for plants comprising a long stemand characteristics of a cauliflower curd. In one embodiment, thepresent invention discloses a method of increasing the stem length of acauliflower plant, for example comprising the steps disclosed above. Thepresent invention also discloses a cauliflower plant obtainable by themethod above, wherein the plant is capable of producing one or moretraits as described herein.

The instant application also provides methods of producing a plant ofthe present invention comprising crossing a cauliflower plant with abroccoli plant and selecting for plants comprising long internodes andcharacteristics of a cauliflower curd. In one embodiment, the presentinvention discloses a method of increasing the stem length of acauliflower plant, for example comprising the steps disclosed above. Inone embodiment, the present invention discloses a method of increasingthe average internode length of the stem of a cauliflower plant, forexample comprising the steps disclosed above. The present invention alsodiscloses a cauliflower plant obtainable by the method above, whereinthe plant is capable of producing one or more traits as describedherein.

The instant application also provides methods of producing a plant ofthe present invention comprising crossing a cauliflower plant with abroccoli plant and selecting for plants comprising a long stem and longinternodes and characteristics of a cauliflower curd. In one embodiment,the present invention discloses a method of increasing the stem lengthof a cauliflower plant, for example comprising the steps disclosedabove. The present invention also discloses a cauliflower plantobtainable by the method above, wherein the plant is capable ofproducing one or more traits as described herein.

In one embodiment, the present invention provides novel cauliflowerplants with a special stem structure. In one embodiment, the novelcauliflower plants provided herein comprise a long stem. In oneembodiment, a cauliflower plant of the present invention is a plant ofthe short cycle or of the medium cycle and comprises at maturity a stemlength of at least about 22.5 cm, optionally at least about 25 cm, inparticular when the plant is grown under conditions representative for acontinental climate, e.g. in Ocsa, Hungary, and planted during the3^(rd) week of May and transplanted during the 3^(rd) week of June, orwhen the plant is grown under conditions representative for aMediterranean climate, e.g. in Murcia, Spain, and planted during the1^(st) week of August and transplanted during the 1st week of September(see e.g. Examples 3 and 4 below). In one aspect, the stem length isabout 22.5 cm to about 35 cm, optionally about 25 cm to about 35 cm, inparticular when grown under the conditions above.

Alternatively, a cauliflower plant of the present invention is a plantof the short cycle or of the medium cycle and comprises at maturity astem length of at least about 30 cm, optionally at least 32.5 cm, whengrown under conditions representative for a continental climate, e.g. inOcsa, Hungary, and planted during the 4^(th) week of May andtransplanted during the 5th week of June (see e.g. Example 5 below). Inone aspect, the stem length is about 30 cm to about 45 cm, optionallyabout 32.5 cm to about 42.5 cm, when grown under the conditions above.

In one embodiment, a cauliflower plant of the present inventioncomprises at maturity a stem length of at least 85% of the length of thestem of a plant of line 03P001 (the male parent of cauliflower hybridCFL5752), representative seeds of which is deposited with NCIMB underaccession number NCIMB 41228. In one aspect, a cauliflower plant of thepresent invention comprises at maturity a stem length of at least 90% ofthe length of the stem of a plant of line 03P001. In one embodiment, acauliflower plant of the present invention comprises at maturity a stemlength of about 85% to about 130%, optionally about 85% to about 120%,about 85% to about 115% or about 85% to about 110%, of the length of thestem of a plant of line 03P001.

In one embodiment, the stem of a plant of the present invention islonger than that of currently existing cauliflower plants havingcommercially desirable attributes, such as capable of producing amarketable curd, and requiring approximately the same number of growingdays to reach maturity, for example at least about 10% longer, at leastabout 15% longer, or at least about 20% longer. Alternatively, the stemof a plant of the present invention is longer than that of currentlyexisting cauliflower plants in the same cycle having commerciallydesirable attributes, such as capable of producing a marketable curd,for example at least about 10% longer, at least about 15% longer, or atleast about 20% longer. In one aspect, the stem of a plant of thepresent invention is about 110% to about 200% the length of the stem ofcurrently existing cauliflower plants having commercially desirableattributes, such as capable of producing a marketable curd, andrequiring approximately the same number of growing days to reachmaturity, optionally about 110% to about 180%. In one aspect, the stemof a plant of the present invention is about 110% to about 200% thelength of the stem of currently existing cauliflower plants in the samecycle having commercially desirable attributes, such as capable ofproducing a marketable curd, optionally about 110% to about 180%.

In one embodiment, such a currently existing cauliflower plants is atypical plants in the cycle. In one embodiment such a currently existingcauliflower plants is a plant of variety Fremont or, alternatively, ofvariety Lecanu and the stem of a plant of the present invention islonger than that of a plant of variety Fremont or variety Lecanu. In oneaspect, the stem of a plant of the present invention is longer than thestem of a plant of variety Fremont or variety Lecanu, in for example atleast about 10% longer, at least about 15% longer, or at least about 20%longer. In one aspect, the stem of a plant of the present invention isabout 110% to about 200% the length of the stem of variety Fremont orvariety Lecanu, optionally about 110% to about 180%.

In one embodiment, the stem of a plant of the present invention reachesa stem length suitable for mechanical harvesting sooner than acauliflower plant of a longer cycle.

In one embodiment, the stem of a plant of the instant inventioncomprises long internodes. In one embodiment, the average internodelength of the stem of a plant of the present invention is at least 1.25cm long (see e.g. Example 5 below). In one aspect, the average internodelength of the stem of a plant of the present invention is at least 1.25cm long when grown under conditions representative for a continentalclimate, e.g. in Ocsa, Hungary, and for example planted during the4^(th) week of May and transplanted during the 5th week of June.Optionally, the average internode length of the stem of a plant of thepresent invention is at least 1.30 cm long, at least 1.35 cm long, or1.40 cm long, especially when grown under the conditions above.Alternatively, the average internode length of the stem of a plant ofthe present invention is 1.25 cm to 2.00 cm, especially when grown underthe conditions above.

In one embodiment, the average internode length of the stem of a plantof the present invention is at least 80% of the average internode lengthof the stem of a plant of line 03P001. In one aspect, the averageinternode length of the stem of a plant of the present invention is atleast 85%, optionally at least 90%, of the average internode length ofthe stem of a plant of line 03P001. In one aspect, the average internodelength of the stem of a plant of the present invention is 80% to 140% ofthe average internode length of the stem of a plant of line 03P001.Optionally, the average internode length of the stem of a plant of thepresent invention is 80% to 130% or 80% to 120% of the average internodelength of the stem of a plant of line 03P001.

In one embodiment, the average internode length of the stem of a plantof the present invention is longer than that of currently existingcauliflower plants having commercially desirable attributes, such ascapable of producing a marketable curd. In one aspect, the averageinternode length of the stem of a plant of the present invention islonger than that of currently existing cauliflower plants havingcommercially desirable attributes, such as capable of producing amarketable curd and requiring approximately the same number of growingdays to reach maturity. Alternatively, the average internode length ofthe stem of a plant of the present invention is longer than that ofcurrently existing cauliflower plants in the same cycle havingcommercially desirable attributes, such as capable of producing amarketable curd.

In one embodiment, the average internode length of the stem of a plantof the present invention is at least 20% longer than the averageinternode length of the stem of a plant of variety Lecanu or,alternatively, of variety Fremont. Optionally, the average internodelength of the stem of a plant of the present invention is at least 25%longer or at least 30% longer than the average internode length of thestem of a plant of variety Lecanu or variety Fremont. Alternatively, theaverage internode length of the stem of a plant of the present inventionis 20% to 80% longer than the average internode length of the stem of aplant of variety Lecanu or variety Fremont.

Typically, cauliflower curds are harvested when they are still tight andcompact, and surrounded by inner leaves. The inner leaves protect thecurd during the harvest by hand. Inner leaves surrounding the curd tendto be associated with large leaves or a large numbers of leaves, orboth. This can cause jams in harvesting machines, and slow downmechanical harvesting of the curds. Accordingly, in one embodiment, aplant of the instant invention further comprises a trait of loosefoliage. In one aspect, the trait of loose foliage is obtained from abroccoli plant.

In one embodiment, the total leaf weight of a plant of the instantinvention is lower than the total leaf weight of currently existingcauliflower plants having commercially desirable attributes, such ascapable of producing a marketable curd. In one aspect, the total leafweight of a plant of the instant invention is lower than the total leafweight of currently existing cauliflower plants having commerciallydesirable attributes, such as capable of producing a marketable curd andrequiring approximately the same number of growing days to reachmaturity. Alternatively, the total leaf weight of a plant of the instantinvention is lower than the total leaf weight of currently existingcauliflower plants in the same cycle having commercially desirableattributes, such as capable of producing a marketable curd.

In one embodiment, the total leaf weight of a plant of the presentinvention is less than 150% of the total leaf weight of a plant of line03P001 (see e.g. Example 5 below). Optionally, the total leaf weight ofa plant of the present invention is less than 140%, less than 130%, lessthan 120 or less than 110%, of the total leaf weight of a plant of line03P001. In one aspect, the total leaf weight of a plant of the presentinvention is 80% to 150% of the total leaf weight of a plant of line03P001.

In one embodiment, the leaf volume index of a plant of the instantinvention is smaller than the leaf volume index of currently existingcauliflower plants having commercially desirable attributes, such ascapable of producing a marketable curd. In one aspect, the leaf volumeindex of a plant of the instant invention is smaller than the leafvolume index of currently existing cauliflower plants havingcommercially desirable attributes, such as capable of producing amarketable curd and requiring approximately the same number of growingdays to reach maturity. Alternatively, the leaf volume index of a plantof the instant invention is smaller than the leaf volume index ofcurrently existing cauliflower plants in the same cycle havingcommercially desirable attributes, such as capable of producing amarketable curd.

In one embodiment, the leaf volume index of a plant of the presentinvention is less than 130% of the leaf volume index of a plant of line03P001 (see Example 5 below). Optionally, the leaf volume index of aplant of the present invention is less than 125%, less than 120%, orless than 110%, of the leaf volume index of a plant of line 03P001. Inone aspect, the leaf volume index of a plant of the present invention is80% to 130% of the leaf volume index of a plant of line 03P001.

In one embodiment, the length of the stem of a plant of the presentinvention is longer because of longer internodes, while the number ofinternodes on the stem, and thus the number of leaves of the plants,remains approximately the same or is reduced, when compared withcurrently existing cauliflower plants having commercially desirableattributes, such as capable of producing a marketable curd and requiringapproximately the same number of growing days to reach maturity or inthe same cycle. The feature of longer internodes thus offers theadvantage of providing longer stems while maintaining a low number ofleaves on the stem or reducing the number of leaves on the stem.

Accordingly, in one embodiment, a plant of the instant inventioncomprises a trait of long stems or long internodes as described herein,and further comprises less than 120% of the number of internodes of aplant of line 03P001. Optionally, a plant of the present inventioncomprises less than 115% or less that 110% of the number of internodesof a plant of line 03P001. In one aspect, plant of the present inventioncomprises 80% to 120% the number of internodes of a plant of line03P001.

In one embodiment, a plant of the present invention comprises 18internodes to 26 internodes. Alternatively, the plant comprises 19internodes to 25 internodes, or 20 internodes to 24 internodes, forexample when the plants are grown under conditions representative for acontinental climate, e.g. in Ocsa, Hungary, and for example plantedduring the 4^(th) week of May and transplanted during the 5th week ofJune.

In one embodiment, a plant of the instant invention further comprises atrait of persistent white curd. A persistent white curd has theadvantage of preventing or slowing down the cauliflower curd turningyellow after exposure to sunlight. The trait of persistent white curd isfor example described in Dickson and Lee (1980)Journal-of-the-American-Society-for-Horticultural-Science, 105(4):533-535 or in Dickson (1985) HortScience, 0(5): 957. In one embodiment,the trait of persistent white is obtained from PI 183214, also describedin the above references. Example 6 below describes the transfer of thepersistent white trait to a cauliflower plant of the instant invention.

Cauliflower plants with a trait of loose foliage produce curds, whichare less protected by inner leaves. Such curds are therefore moreexposed to sunlight. Accordingly, in one embodiment, the trait ofpersistent white is combined with the trait of loose foliage, and in oneembodiment, a plant of the instant invention further comprises a traitof loose foliage and a trait of persistent white curd. Such cauliflowerplants have the advantage of producing fewer or smaller leaves, and thusprevent jamming of mechanical harvesters, while preventing or slowingdown the cauliflower curd turning yellow after exposure to sunlight.

When plants are compared in the context of the instant application, forexample when a trait is compared between a plant of the presentinvention and a plant of line 03P001 or a plant of variety Fremont orvariety Lecanu, the plants are typically grown under the sameconditions, for example in the same experiment. In one aspect, theplants are grown under the conditions set forth in the examples below,for example under conditions representative for a continental climate,e.g. in Ocsa, Hungary, and for example planted during the 4^(th) week ofMay and transplanted during the 5th week of June.

In one embodiment, a plant of the present invention comprises anycombination the traits of long stem, long internodes, loose foliage,number of internodes, total leaf weight, leaf volume index andpersistent white curd, and short or medium cycle and uniformity, asdescribed herein. In one embodiment, a plant of the present inventioncomprises a long stem, loose foliage and persistent white curd, asdescribed herein. In one embodiment, such plant is of the short cycle ormedium cycle.

In one embodiment, a plant of the present invention comprises a longstem and long internodes, as described herein. In one aspect, the plantcomprises a long stem, long internodes and number of internodes, asdescribed herein. In another aspect, the plant comprises a long stem,long internodes and low total leaf weight, as described herein.Alternatively, the plant comprises a long stem, long internodes andsmall leaf volume index, as described herein. Optionally, the plantcomprises a long stem, long internodes, number of internodes, low totalleaf weight and small leaf volume index, as described herein. In oneembodiment, the plant is of the short cycle or medium cycle.

In one embodiment, a plant of the present invention comprises a longstem and low total leaf weight, as described herein. In one aspect, theplant comprises a long stem, long internodes and number of internodes,as described herein. In another aspect, the plant comprises a long stemand small leaf volume index, as described herein. Alternatively, theplant comprises a long stem, number of internodes, low total leaf weightand small leaf volume index, as described herein. In one embodiment, theplant is of the short cycle or medium cycle.

In one embodiment, a plant of the present invention comprises longinternodes and low total leaf weight, as described herein. In oneaspect, the plant comprises long internodes and number of internodes, asdescribed herein. In another aspect, the plant comprises long internodesand small leaf volume index, as described herein. Alternatively, theplant comprises long internodes, number of internodes, low total leafweight and small leaf volume index, as described herein. In oneembodiment, the plant is of the short cycle or medium cycle.

In one embodiment, plants of the instant invention of a line or hybridhave a high level of uniformity in plant habit. In one embodiment,plants of the instant invention of a line or hybrid have a high level ofuniformity in growing days. In one embodiment, plants of the instantinvention of a line or hybrid have a high level of uniformity in planthabit and growing days. These features facilitate the use of fully orpartly automatic mechanical harvesters. High level of uniformity inplant type is desirable for high through-put machine harvest to avoidlosses due to the different plant shapes, while uniformity in maturationlimits the number of passes over the field required, further decreasingthe cost of the harvest. This results in a high proportion of marketableproducts. High uniformity makes frequent adaptation of the harvester, inextreme case one-by-one examination of plants, unnecessary during theharvest process and allows use of a machine at its highest capacity. Inone embodiment, high level of uniformity is achieved by the applicationof double haploid technique on the plants of the instant invention (seee.g. Bagga, S. et al. (1982). Comparison of in vitro plant formationfrom somatic tissues and pollen grains in Brassica oleracea var.botrytis. Phytomorphology 32, p 152-156; Ockendon, D. J. (1988). Theploidy of plants obtained from anther culture of cauliflowers (Brassicaoleracea var. botrytis). Ann. appl. Biol. 113, p 319-325). Accordingly,in one embodiment, a plant of the present invention is a dihaploid orthe descendent of a dihaploid plant, for example a hybrid having adihaploid as parent line.

Based on the description of the present invention, the skilled person isable to recognize a cauliflower plant of the instant invention undervarious growing conditions. Accordingly, the present invention alsofurther discloses a method of identifying a cauliflower plant of theinstant invention comprising growing a cauliflower plant until saidplant reaches maturity and measuring the length of the stem, length ofinternodes, total leaf weight or leaf volume index of said plant. Basedon the description of the present invention, the skilled person is alsoable to transfer a trait of cauliflowers plant of the present inventionto other cauliflower plants, in particular to any type of cauliflowertypes using breeding techniques well-known in the art. Cauliflower typesinclude for example white cauliflower, green cauliflower, purplecauliflower and orange cauliflower, or cauliflower of the Romano type.Based on the description of the present invention, the skilled person isalso able to transfer a trait of cauliflowers plant of the presentinvention to cauliflower plants of other cycles using breedingtechniques well-known in the art.

The values for stem length, average internode length, number ofinternodes, total leaf weight, leaf length, leaf width and leaf volumeindex disclosed in the instant application are averages based themeasurement of these characteristics for a representative number ofplants, unless otherwise stated.

Representative cauliflower plants according to the instant invention aredescribed in the Examples below. For example, cauliflower hybrid CFL5752of the present invention (medium cycle) was grown in the same trial asvariety Fremont (medium cycle), variety Lecanu (medium cycle) andvariety Amerigo (medium-long cycle). The stem length was measured atmaturity and is reported in Table 1. Generally, the stem length isreported herein as an average, typically calculated from the measurementof 15 to 20 stems of plants of a line or hybrid. It is understood thatmeasurements in independent experiments may vary because of differentenvironmental conditions.

Seeds of line 03P001, a representative line according to the instantinvention, were deposited under the Budapest treaty with NCIMB, Ltd.,Aberdeen, Scotland, UK, on Jun. 28, 2004 under accession number NCIMB41228. Line 03P001 is the male parent of cauliflower hybrid CFL5752 andis also further described in the Examples herein.

In one embodiment, a plant of the present invention is obtainable from aplant of line 03P001, seeds of which was deposited under accession no.NCIMB 41228, .or from a progeny or ancestor of said line, whichcomprises long stems according to the instant invention.

In one embodiment, the present invention further discloses a method oftransferring one or more traits according to the present invention, forexample long stems, to a cauliflower plant lacking said trait(s)comprising a) obtaining a plant comprising said trait(s); b) crossing itto a plant lacking said trait(s); c) obtaining plants of the cross ofstep b); d) selecting a plant of step c) which is capable of producingone or more traits according to the present invention. Optionally, themethod further comprises e) back-crossing a plant resulting from step d)with a cauliflower plant, and f) selecting for a cauliflower plant,which is capable of producing one or more traits according to thepresent invention. In one aspect, the method further comprises obtainingan inbred cauliflower plant, which is capable of producing one or moretraits according to the present invention, and, in another aspect, themethod further comprises crossing said inbred cauliflower plant toanother cauliflower plant to produce a hybrid cauliflower plant, whichis capable of producing one or more traits according to the presentinvention. In one embodiment, the plant of step a) comprising saidtrait(s) is a plant of line 03P001 or a progeny or ancestor of saidplant. In one embodiment, the plant of step a) is a broccoli plant.

In one embodiment, the present invention discloses a cauliflower plantobtainable by any one of the methods above, wherein the plant is capableof producing one or more trait as described herein.

In one embodiment, the present invention discloses a method of producinga plant comprising one or more traits according to the presentinvention, for example long stems, to a cauliflower plant lacking saidtrait(s) comprising a) obtaining a plant comprising said trait(s); b)crossing it to a plant lacking said trait(s); c) obtaining plants of thecross of step b); d) selecting a plant of step c) which is capable ofproducing one or more traits according to the present invention.Optionally, the method further comprises e) back-crossing a plantresulting from step d) with a cauliflower plant, and f) selecting for acauliflower plant, which is capable of producing one or more traitsaccording to the present invention. In one aspect, the method furthercomprises obtaining an inbred cauliflower plant, which is capable ofproducing one or more traits according to the present invention, and, inanother aspect, the method further comprises crossing said inbredcauliflower plant to another cauliflower plant to produce a hybridcauliflower plant, which is capable of producing one or more traitsaccording to the present invention. In one embodiment, the plant of stepa) comprising said trait(s) is a plant of line 03P001 or a progeny orancestor of said plant. In one embodiment, the plant of step a) is abroccoli plant.

In one embodiment, the present invention discloses a cauliflower plantobtainable by any one of the methods above, wherein the plant is capableof producing one or more traits as described herein.

Commercial cauliflowers are generally hybrids produced from the cross oftwo parental lines (inbreds). The development of hybrids requires, ingeneral, the development of homozygous inbred lines, the crossing ofthese lines, and the evaluation of the crosses. Pedigree breeding andrecurrent selection breeding methods are used to develop inbred linesfrom breeding populations. Breeding programs combine the geneticbackgrounds from two or more inbred lines or various other germplasmsources into breeding pools from which new inbred lines are developed byselfing and selection of desired phenotypes. The new inbreds are crossedwith other inbred lines and the hybrids from these crosses are evaluatedto determine which of those have commercial potential. Plant breedingand hybrid development are expensive and time-consuming processes.

Pedigree breeding starts with the crossing of two genotypes, each ofwhich may have one or more desirable characteristics that is lacking inthe other or which complements the other. If the two original parents donot provide all the desired characteristics, other sources can beincluded in the breeding population. In the pedigree method, superiorplants are selfed and selected in successive generations. In thesucceeding generations the heterozygous condition gives way tohomogeneous lines as a result of self-pollination and selection.Typically in the pedigree method of breeding five or more generations ofselfing and selection is practiced: F1 to F2; F3 to F4; F4 to F5, etc.

A single cross hybrid results from the cross of two inbred lines, eachof which has a genotype that complements the genotype of the other. Thehybrid progeny of the first generation is designated F1. In thedevelopment of commercial hybrids only the F1 hybrid plants are sought.Preferred F1 hybrids are more vigorous than their inbred parents. Thishybrid vigor, or heterosis, can be manifested in many polygenic traits,including increased vegetative growth and increased yield.

Cauliflower plants can be easily cross-pollinated. A trait is readilytransferred from one cauliflower plant to another cauliflower plant,including cauliflower plants of different types using conventionalbreeding techniques, for example to further obtain commercial lines. Theintrogression of a trait into the elite line is for example achieved byrecurrent selection breeding, for example by backcrossing. In this case,the elite line (recurrent parent) is first crossed to a donor inbred(the non-recurrent parent) that carries the trait. The progeny of thiscross is then mated back to the recurrent parent followed by selectionin the resultant progeny for the trait. After three, preferably four,more preferably five or more generations of backcrosses with therecurrent parent with selection for the trait, the progeny isheterozygous for the locus harboring the resistance, but is like therecurrent parent for most or almost all other genes (see, for example,Poehlman & Sleper (1995) Breeding Field Crops, 4th Ed., 172-175; Fehr(1987) Principles of Cultivar Development, Vol. 1: Theory and Technique,360-376, incorporated herein by reference). Selection for the trait iscarried out after each cross.

In one embodiment, a cauliflower plant of the present invention is malesterile. Male sterility is of value in B. oleracea hybrid seed breedingbecause normal flowers are self-pollinating. Male sterile lines do notproduce viable pollen and cannot self-pollinate. By eliminating thepollen of one parental variety in a cross, a plant breeder is assured ofobtaining hybrid seed of uniform quality. A particularly advantageousmale sterility system is cytoplasmic male sterility (CMS). An example ofsuch CMS in Brassica is the Ogura CMS originally found in radish (seefor example Ogura (1968) Mem. Fac. Agric. Kagoshima Univ. 6: 39-78;Makaroff (1989) Journal of Biol. Chem. 264: 11706-11713; U.S. Pat. No.5,254,802). Therefore, the present invention discloses a male sterile,in particular CMS, cauliflower plant, including seeds and materials ofsaid plants and the progeny thereof. In one embodiment, a plant of theinstant invention is a maintainer plant.

The male fertility of male sterile plants can be restored by methodswell-known in the art. The male fertility of CMS plants, in particularCMS B. oleracea plants, is preferably restored by cell fusion. For this,cells of a CMS plant are fused to cells of a male fertile plant toreplace the nucleus of the fertile plant by the nucleus of sterile plantin the fertile cytoplasmic background, and restore fertility. Cellfusion techniques are well-known in the art and are for exampledescribed in Sigareva and Earle (1997) Theor. Appl. Genet. 94: 213-320.Using such techniques, male fertile plants are regenerated, and allowedto self-pollinate or crossed to another plant.

In one embodiment, a plant of the present invention is an inbred, ahybrid, or a dihaploid, for example produced by pedigree breeding or byrecurrent selection breeding. In one embodiment, a plant of the presentinvention has commercially acceptable agronomic characteristics.

In one embodiment, the present invention discloses a method of producingseed of a cauliflower plant of the present invention comprising: a)growing a plant of the present invention; b) allowing said plant toself-pollinate; c) harvesting seeds from said plant.

Cauliflower plants can also be propagated vegetatively using methodswell-known in the art, for example in-vitro plant tissue culture,rooting side shoot or protoplast fusion. In one embodiment, a method ofvegetatively propagating a plant of the present invention comprises: a)collecting tissue of a plant of the present invention; b) cultivatingsaid tissue to obtain proliferated shoots; c) rooting said proliferatedshoots to obtain rooted plantlets; d) growing plants from said rootedplantlets; and harvesting seeds from said plants.

Cauliflower plants of the present invention can also be transformedgenetically with a gene of interest, using techniques well known in theart. Accordingly, the present invention also further disclose acauliflower plant according to the instant invention further such a geneof interest.

In one embodiment, the present application discloses methods ofproducing a cauliflower curd comprising growing a plant of the inventionuntil a curd is produced and harvesting the curd. In one aspect, thecurd is harvested mechanically.

In one embodiment, the present application discloses methods ofharvesting a cauliflower curd comprising growing a plant of theinvention until a curd is produced and mechanically harvesting saidcurd. In one aspect, a blade is used in the step of mechanicallyharvesting the curd. In one embodiment, the curd is loaded on a belt.

All references cited herein are incorporated by reference in the instantapplication in their entireties. The following examples are intended toprovide illustrations of the application of the present invention. Thefollowing examples are not intended to completely define or otherwiselimit the scope of the invention.

EXAMPLES Example 1 Development of Line 03P001 and of Hybrid CFL5752

Plants of cauliflower line B66-1, a Syngenta breeding line, were crossedto plants of a Syngenta broccoli breeding line. The resulting F1 progenywas self-pollinated and selected for long stem and internode, and forthe best possible cauliflower curd. Selected F2 progenies were againself-pollinated and selected for long stem and internode and for thebest possible cauliflower curd. These steps were repeated for selectedF3 progenies.

Selected F3 progenies were subjected to double haploid culture using astandard protocol. The resulting dihaploid lines were again selected forlong stem and internode and for the best possible cauliflower curd. Line03P001 was identified.

About 20 lines were crossed with the female parent of hybrid commercialhybrid Amerigo (Syngenta Seeds) to obtain F1 hybrids. In particular,line 03P001 was crossed with the female parent of Amerigo to obtainhybrid CFL5752.

Example 2 Characteristics of Hybrid CFL5752

Hybrid CFL 5752 typically reaches maturity at approximately 63 daysafter transplanting, when seeds are sown in January (week 3) and youngplants transplanted in week 13, under growing conditions in Enkhuizen,The Netherlands.

Example 3 Measurements of Stem Length

The stem length of cauliflower hybrid CFL5752 was compared to that ofvarieties Fremont and Amerigo, and to that of the female parent ofAmerigo and CFL5752. Variety Fremont is a medium cycle variety ofSeminis. Amerigo is a medium-long cycle variety of Syngenta. Fremont andAmerigo were used standard varieties for the medium cycle and themedium-long cycle, respectively.

Experiment 1 was conducted in Ocsa, Hungary, under conditionsrepresentative for a continental climate. In Experiment 1, seeds weresown on May 18, 2004 and transplanted on Jun. 16, 2004. Plants weregrown in open fields watered by sprinklers, with a plant density ofapproximately 3 plant/m² (60 cm row distance, 55 cm plant distancewithin a row). The average maximal temperature during the growing periodwas: June 24.1° C.; July 26.6° C.; August 26.9° C. The average minimumtemperature during the growing period was: June 13.8° C.; July 16.7° C.;August 15.4° C.

Experiment 2 was conducted in Murcia, Spain, under conditionsrepresentative for a mediteranean climate. In Experiment 2, seeds weresown on Aug. 2, 2004 and transplanted on Sep. 3, 2004 Plants were grownin open field with drip irrigation, with a plant density ofapproximately 3,3 plant/m² (1 m row distance, 30 cm plant distancewithin a row). The average maximal temperature during the growing periodwas: Sept. 27.0° C.; Oct. 24.2° C.; Nov. 17.9° C.; Dec. 15.7° C. Theaverage minimum temperature during the growing period was: Sept. 18.5°C.; Oct. 13.3° C.; Nov. 7.3° C.; Dec. 9.0° C.

Plants were grown until the curds of 50% of the plants of a certain plotwere ready to be harvested. This time point was calculated from the dateyoung plants were transplanted and is reported in the Tables as thenumber of growing days in the field. At this time point, plants havingachieved an optimal harvest time were selected and measurements of thelength of their stems was carried out. The number of growing days in thesecond trial is higher than in the first trial. This is because thenight temperature was higher in Murcia (Exp. 2) than in Ocsa (Exp. 1).Table 1 shows the number of stems measured, the average stem length andthe standard deviation.

Table 2 shows the results for individual plots in Experiment 2, leadingto the averages shown in Table 1. TABLE 1 Exp. 1 (2004) Exp. 2 (2004)Harvest stem stem time length days after length days after (week (cm)transplanting (cm) transplanting number) CFL5752 47-50 Avg 26.3 56 28.384.6 Std Dev 3.0 2.7 No. of plants 27 94 Fremont 47-49 Avg 19.8 56 18.980.4 Std Dev 1.9 2.5 No. of plants 40 78 Amerigo 51-52 Avg 28.3 70 28.0104.4 Std Dev 2.1 1.5 No. of plants 40 87 Female of 47-50 Amerigo Avg28.9 61 26.7 89 Std Dev 2.9 3.1 No. of plants 18 19

TABLE 2 Individual plots in Exp. 2: stem length (cm) days aftertransplanting, avg. over plots CFL5752 2005/2 2005/32 2005/48 2005/692005/104 84.6 Avg 26.1 30.3 28.6 28.8 27.5 Std Dev 2 1.9 3.4 2.2 2.1 Noof plants 18 19 20 17 20 Fremont 2005/3 2005/33 2005/49 2005/70 80.4 Avg19.0 19.5 17.4 19.3 Std Dev 2.8 1.7 2.1 3.1 No of plants 20 20 18 20Amerigo 2005/1 2005/31 2005/47 2005/68 2005/103 104.4 Avg 26.3 28.9 27.828.3 28.6 Std Dev 0.8 1.2 1.6 1.4 1.0 No of plants 17 18 18 17 17 Femaleof Amerigo  2005/50 89 Avg 26.7 Std Dev 3.1 No of plants 19

Example 4 Measurements of Stem Length

A number of representative cauliflower plants of the instant inventionare described below. Plants were grown under the same conditions are inExperiment 2 in Example 3 above. The stem length was measured and shownin Table 3 below.

Female of Amerijo: Medium vigour, erected plant habit, medium stemlength and internode length. Plant is open, curd shape is flat, curddensity is medium. Medium-long cycle.

Male of CFL5752 (line 03P001): Medium vigour, erected plant habit, longstem and internodes. Plant is open, curd shape is domic and free ofharriness, curd density is good. Medium cycle.

Handcross 14: Vigorous hybrid plant with long stem and long internodes.Plant is open, curd shape is domic, smooth and free of harriness. Curddensity is medium. Medium cycle.

Handcross 17: Medium vigour, erected hybrid plant habit, long stem andinternodes. Plant is open, curd shape is domic and free of harriness.Curd density is medium. Medium cycle.

Handcross 64: Hybrid plant with medium vigour, long stem and internodes.Plant is open, curd shape is domic and free of harriness. Curd densityis medium. Medium cycle.

F6 line: Vigorous plant with long stem and long internodes and openbottom closure.

Plant is open, curd shape is flat, smooth and free from harriness. Curddensity is modest. Medium cycle.

DH line 122: Dihaploid with medium vigour, long stem and internodes andopen bottom closure. Plant is open, curd shape is flat and free ofharriness. Curd density is modest. Medium cycle.

DH line 123: Dihaploid with medium vigour, long stem and internode.Plant is open, curd shape is flat and free of harriness. Curd density ismodest. Medium cycle. TABLE 3 Harvest Harvest stem days time stem daystime length after (week length after (week (cm) transplanting number)(cm) transplanting number) Female 2005/50 89 47-50 DH line (male 2005/5489 48-50 of Amerigo of CFL5752) Avg 26.7 Avg 30.2 Std Dev 3.1 Std Dev1.6 No. of plants 19 No. of plants 19 Handcross 14 2005/14 78 47-50Handcross 17 2005/17 77 47-49 Avg 30.3 Avg 30.4 Std Dev 2.4 Std Dev 2.5No. of plants 16 No. of plants 18 Handcross 64 2005/64 78 47-49 line F62005/90 87 47-50 Avg 28.6 Avg 28.0 Std Dev 2.9 Std Dev 3.5 No. of plants20 No. of plants 16 DH line 122  2005/122 71 47-48 DH line 123  2005/12375 47-48 Avg 32.7 Avg 31.0 Std Dev 2.1 Std Dev 1.8 No. of plants 20 No.of plants 20

Example 5 Comparative Measurements

The stem length, number of leaves, average internode length, total leafweight, leaf length, leaf width and leaf volume of a number ofrepresentative plants of the present invention are described in Table 4.Variety Amerigo was used as a standard variety for the medium-longcycle. Varieties Fremont and Lecanu were used a standard varieties forthe medium cycle.

The experiment was conducted in Ocsa, Hungary, under conditionsrepresentative for a continental climate. Seeds were sown on may 27,2005 and transplanted on Jun. 28, 2005. Plants were grown in open fieldswatered by sprinklers, with a plant density of approximately 3 plant/m²(60 cm row distance, 55 cm plant distance within a row). The averagemaximal temperature during the growing period was: July 23.6° C.; August26.9° C.; September 24.6° C. The average minimum temperature during thegrowing period was: July 15.7° C.; August 15.4° C.; September 12.3° C.

Plants were grown until the curds of 50% of the plants of a certain plotwere ready to be harvested. This time point was calculated from the dateyoung plants were transplanted and is reported in the Tables as thenumber of growing days in the field (days after transplanting). At thistime point, plants having achieved an optimal harvest time were selectedand measurements were carried out when individual plants were ready forharvest.

Table 4 shows the number of plant for each entry and the average andstandard deviation for each measurement. For the measurements of leaflength and width, 3 fully developed leaves were measured for eachindividual plant.

The number of growing days in the second trial in Ocsa was higher thanin the first trial at this location (Experiment 1, Table 1). The secondtrial was planted two weeks later than the first one. The temperaturewas warmer when the plants needed vernalization for curd induction. Thiscaused approximately two weeks delay in maturity and had an effect onstem length, leading to longer stems in the second experiment. TABLE 4Leaf volume Maturity Stem length Number of Internode Total leaf Leaflength Leaf width index = length × (days after (cm) internodes length(cm) weight (kg) (cm) (cm) width/1000 planting) Amerigo 83.1 Average34.8 29.9 1.16 1.625 54.07 23.45 1.27 Std Dev 2.6 1.8 0.13 0.273 4.232.42 0.19 No. plants: 14 CFL5752 73.4 Average 35.2 23.4 1.50 1.154 55.4424.19 1.34 Std Dev 3.3 3.2 0.26 0.273 3.71 2.40 0.21 No. plants: 27Fremont 69.9 Average 26.2 23.6 1.11 1.363 62.84 25.23 1.59 Std Dev 2.53.1 0.11 0.211 7.12 2.40 0.29 No. plants: 19 Lecanu 74.4 Average 26.926.0 1.03 1.481 58.05 26.08 1.51 Std Dev 2.7 2.2 0.13 0.350 4.84 4.140.35 No. plants: 26 Female of 83.3 Amerigo (fertile) Average 33.8 28.31.19 1.267 51.96 22.74 1.18 Std Dev 2.0 3.2 0.20 0.139 2.88 1.90 0.15No. plants: 9 Female of 85.0 Amerigo (CMS) Average 31.3 27.8 1.12 1.40451.75 23.31 1.21 Std Dev 3.1 4.1 0.17 0.405 4.50 3.50 0.29 No. plants:12 Male of CFL5752 73.6 Average 33.7 22.2 1.52 0.798 48.25 22.77 1.10Std Dev 2.5 2.2 0.20 0.252 4.94 2.99 0.26 No. plants: 20 Handcross 6471.6 Average 31.1 21.1 1.48 1.050 54.73 24.80 1.36 Std Dev 4.0 3.9 0.320.338 7.55 3.50 0.34 No. plants: 18 Handcross 14 76.9 Average 35.1 23.01.53 1.218 53.53 22.47 1.20 Std Dev 4.6 3.1 0.26 0.340 6.33 2.51 0.26No. plants: 20 New DH line 123 71.0 Average 36.1 21.8 1.66 0.810 51.8321.22 1.10 Std Dev 5.5 3.1 0.37 0.3 4.7 1.8 0.2 No. plants: 24

Example 6 Production of a Cauliflower Plant Comprising the Trait ofPersistent White Curd

The original source of the trait of persistent white curd was anEgyptian landrace derived from Genebank PI 183214. Besides the persitentwhiteness, PI 183214 carries many undesirable traits, such as poor curdquality and susceptibility for downy mildew. The original persistentwhite material was crossed with plants of a cauliflower elite line withgood curd quality. The resulting F1 progeny was self-pollinated and theresulting progeny was planted in open fields. The best plants wereself-pollinated again. The persitent white plants with the best curdquality and plant habit were selected in the F3 population. These plantswere used as a donor of the trait of persistent white curd with bettercurd quality and plant habit than the original source.

These plants were crossed again with cauliflower plants to furtherimprove plant habit and curd quality. Elite lines were developed from F2populations derived from these crosses. Plants of broccoli lines werecrossed with some of these F2 plants to introduce the trait of longstem, long internode and loose foliage in these plants. Before startingline development, one backcross with plants of a cauliflower linecomprising the trait of persistent white curd was performed. Linefixation was carried out using the dihaploid technique.

1. A method for producing a cauliflower plant with a long stemcomprising: a) crossing a broccoli plant and a cauliflower plant; b)obtaining a progeny plant from the cross of step a); c) selecting aprogeny plant comprising at maturity a stem with an average internodelength of at least 1.25 cm.
 2. The method according to claim 1,comprising selecting a progeny plant comprising at maturity a stem withan average internode length of at least 1.30 cm.
 3. The method accordingto claim 1, further comprising self-pollinating said progeny plantselected in step c).
 4. The method according to claim 1, furthercomprising obtaining a dihaploid of said progeny plant selected in stepc).
 5. A cauliflower plant obtained by the method of claim 1, whereinsaid plant comprises at maturity a stem with an average internode lengthof at least 1.25 cm.
 6. The cauliflower plant according to claim 5,wherein said plant comprises at maturity a stem with an averageinternode length of at least 1.30 cm.
 7. The method according to claim1, wherein said plant comprises at maturity a stem with an averageinternode length of 1.25 cm to 2.00 cm.
 8. The cauliflower plantaccording to claim 5, wherein said stem comprises 18 internodes to 26internodes.
 9. The cauliflower plant according to claim 5, wherein thelength of the stem of said plant is at least 85% of the length of thestem of a plant of line 03P001, representative seed of which beingdeposited under accession number NCIMB
 41228. 10. The cauliflower plantaccording to claim 5, wherein said plant is a plant of the short cycleor of the medium cycle.
 11. The cauliflower plant according to claim 5,wherein the total leaf weight of said plant is less than 150% of thetotal leaf weight of a plant of line 03P001, representative seed ofwhich being deposited under accession number NCIMB
 41228. 12. Thecauliflower plant according to claim 5, wherein the leaf volume index ofsaid plant is less than 130% of the leaf volume index of a plant of line03P001, representative seed of which being deposited under accessionnumber NCIMB
 41228. 13. The cauliflower plant according to claim 5,wherein said plant is an inbred line, a hybrid or a dihaploid.
 14. Thecauliflower plant according to claim 5, wherein said plant ismale-sterile.
 15. Seed of a cauliflower plant according to claim
 5. 16.Part of a cauliflower plant according to claim
 5. 17. Part according toclaim 16, wherein said part is a cell, pollen, an ovule, or a curd. 18.A method of producing a cauliflower curd comprising growing a plantaccording to claim 5 until a curd is produced and harvesting said curd.19. The method according to claim 18, wherein said curd is harvestedmechanically.
 20. A method of harvesting a cauliflower curd comprisinggrowing a plant according to claim 5 until a curd is produced andmechanically harvesting said curd.
 21. The method according to claim 20,wherein a blade is used in said step of mechanically harvesting saidcurd.
 22. A method of producing seed of a cauliflower plant comprising:a) growing a cauliflower plant according to claim 5; b) allowing saidcauliflower plant to self-pollinate; and c) harvesting seeds from saidcauliflower plant.
 23. A method of vegetatively propagating acauliflower plant comprising: a) collecting a tissue of a cauliflowerplant according to claim 5; b) cultivating said tissue to obtainproliferated shoots; c) rooting said proliferated shoots to obtainrooted plantlets.
 24. A method for producing a cauliflower plant: a)crossing a cauliflower plant and a plant of line 03P001, representativeseed of which being deposited under accession number NCIMB 41228; b)obtaining a progeny plant from the cross of step a); c) selecting aplant comprising at maturity a stem with an average internode length ofat least 1.25 cm.
 25. The method according to claim 24, furthercomprising obtaining a dihaploid of said progeny plant selected in stepc).
 26. Seed of a plant of line 03P001, representative seed of whichbeing deposited under accession number NCIMB
 41228. 27. Plant grown froma seed of claim
 26. 28. Part of a plant of claim
 26. 29. A cauliflowerplant comprising at maturity a stem with a length of at least 85% of thelength of the stem of a plant of line 03P001, representative seed ofwhich being deposited under accession number NCIMB 41228, wherein saidplant is a plant of the short cycle or of the medium cycle.
 30. Thecauliflower plant according to claim 29, wherein the length of said stemis at least 90% of the length of a stem of a plant of said line 03P001.31. The cauliflower plant according to claim 29, wherein the length ofsaid stem is 85% to 130% of the length of a stem of a plant of said line03P001.
 32. The cauliflower plant according to claim 29, wherein thenumber of internodes of said plant is less than 120% of the number ofinternodes of a plant of said line 03P001.
 33. The cauliflower plantaccording to claim 29, wherein the total leaf weight of said plant isless than 150% of the total leaf weight of a plant of line 03P001,representative seed of which being deposited under accession numberNCIMB
 41228. 34. The cauliflower plant according to claim 29, whereinthe leaf volume index of said plant is less than 130% of the leaf volumeindex of a plant of line 03P001, representative seed of which beingdeposited under accession number NCIMB
 41228. 35. Seed of a plantaccording to claim
 29. 36. Part of a plant according to claim 29.